Bacteriophages are currently used as an alternative method for controlling bacterial spot disease on tomato incited by Xanthomonas campestris pv. vesicatoria. However, the efficacy of phage is greatly reduced due to its short residual activity on plant foliage. Three formulations that significantly increased phage longevity on the plant surface were tested in field and greenhouse trials: (i) PCF, 0.5% pregelatinized corn flour (PCF) + 0.5% sucrose; (ii) Casecrete, 0.5% Casecrete NH-400 + 0.5% sucrose + 0.25% PCF; and (iii) skim milk, 0.75% powdered skim milk + 0.5% sucrose. In greenhouse experiments, the nonformulated, PCF-, Casecrete-, and skim milk-formulated phage mixtures reduced disease severity on plants compared with the control by 1, 30, 51, and 62%, respectively. In three consecutive field trials, nonformulated phage caused 15, 20, and 9% reduction in disease on treated plants compared with untreated control plants, whereas plants treated with PCF- and Casecrete-formulated phage had 27, 32, and 12% and 30, 43, and 24% disease reduction, respectively. Plants receiving copper-mancozeb treatments were included in two field trials and had a 20% decrease in disease in the first trial and a 13% increase in the second one. Skim milk-formulated phage was tested only once and caused an 18% disease reduction. PCF-formulated phage was more effective when applied in the evening than in the morning, reducing disease on plants by 27 and 13%, respectively. The Casecrete-formulated phage populations were over 1,000-fold higher than the nonformulated phage populations 36 h after phage application.
Various combinations of the harpin protein, acibenzolar-S-methyl, and bacteriophages were compared for controlling tomato bacterial spot in field experiments. Harpin protein and aciben-zolar-S-methyl were applied every 14 days beginning twice before transplanting and then an additional four applications throughout the season. Formulated bacteriophages were applied prior to inoculation followed by twice a week at dusk. A standard bactericide treatment, consisting of copper hydroxide plus mancozeb, was applied once prior to inoculation and then every 7 days, while untreated plants served as an untreated control. Experiments were conducted in north and central Florida fields during fall 2001, spring 2002, and fall 2002. In three consecutive seasons, acibenzolar-S-methyl applied in combination with bacteriophage or bacteriophage and harpin significantly reduced bacterial spot compared with the other treatments. However, it did not significantly affect the total yield compared with the standard or untreated control. Application of host-specific bacteriophages was effective against the bacterial spot pathogen in all three experiments, providing better disease control than copper-mancozeb or untreated control. When results of the disease severity assessments or harvested yield from the bacteriophage-treated plots were grouped and compared with the results of the corresponding nonbacteriophage group, the former provided significantly better disease control and yield of total marketable fruit.
Two strains of plant growth-promoting rhizobacteria, two systemic acquired resistance inducers (harpin and acibenzolar-S-methyl), host-specific unformulated bacteriophages, and two antagonistic bacteria were evaluated for control of tomato bacterial spot incited by Xanthomonas campestris pv. vesicatoria in greenhouse experiments. Untreated plants and plants treated with copper hydroxide were used as controls. The plant growth-promoting rhizobacteria or a tap water control were applied as a drench to the potting mix containing the seedlings, while the other treatments were applied to the foliage using a handheld sprayer. The plant growth-promoting rhizobacteria strains, when applied alone or in combination with other treatments, had no significant effect on bacterial spot intensity. Messenger and the antagonistic bacterial strains, when applied alone, had negligible effects on disease intensity. Unformulated phage or copper bactericide applications were inconsistent in performance under greenhouse conditions against bacterial spot. Although acibenzolar-S-methyl completely prevented occurrence of typical symptoms of the disease, necrotic spots typical of a hypersensitive reaction (HR) were observed on plants treated with acibenzolar-S-methyl alone. Electrolyte leakage and population dynamics experiments confirmed that acibenzolar-S-methyl-treated plants responded to inoculation by eliciting an HR. Application of bacteriophages in combination with acibenzolar-S-methyl suppressed a visible HR and provided excellent disease control. Although we were unable to quantify populations of the bacterium on the leaf surface, indirectly we determined that bacteriophages specific to the target bacterium reduced populations of a tomato race 3 strain of the pathogen on the leaf surface of acibenzolar-S-methyl-treated plants to levels that did not induce a visible HR. Integrated use of acibenzolar-S-methyl and phages may complement each other as an alternative management strategy against bacterial spot on tomato.
Greenhouse experiments were conducted to determine the effectiveness of plant essential oils as soil fumigants to manage bacterial wilt (caused by Ralstonia solanacearum) in tomato. Potting mixture (“soil”) infested with R. solanacearum was treated with the essential oils at 400 mg or μl and 700 mg or μl per liter of soil in greenhouse experiments. R. solanacearum population densities were determined just before and 7 days after treatment. Populations declined to undetectable levels in thymol, palmarosa oil, and lemongrass oil treatments at both concentrations, whereas tea tree oil had no effect. Tomato seedlings transplanted in soil treated with 700 mg/liter of thymol, 700 ml/liter of palmarosa oil, and 700 ml/liter of lemongrass oil were free from bacterial wilt and 100% of plants in thymol treatments were free of R. solanacearum. Soil amendment with fresh leaves of essential oil-producing plants did not reduce bacterial wilt incidence compare to untreated inoculated control. Some thyme oil-producing plants such as thyme (Thymus vulgaris) cv. German winter, Creeping thyme (Thymus serpyllum), and Greek oregano (Origanum vulgare subsp. hirtum), while remaining symptomless, became systemically infected by R. solanacearum and were therefore identified as hosts of R. solanacearum.
There are few growth studies evaluating within‐season effects of N on vegetative growth and N accumulation of tomato (Lycopersicon esculentum Mill.). Growth analysis of field‐grown tomato for a number of Florida locations and management systems is presented here. Severe N stress resulted in fewer and smaller, but thicker, leaves. With increasing N, average leaf area index (LAI) increased from ≈0.75 to ≈3, but radiation use efficiency (RUE) typically increased less then 30%. Lower RUE under N‐limited conditions reflected a decrease in N concentration of the most recently matured leaves from 40 mg g−1 to as little as 15 mg g−1. Over the life of well‐fertilized crops, leaf N concentrations dropped from 55 to 65 mg g−1 during initial growth to 20 to 35 mg g−1 at final harvest. Corresponding N concentrations for fruit and for stems were 30 to 35 mg g−1 and 15 to 25 mg g−1. Severe N stress affected leaf and stem N concentrations most drastically, whereas N in fruits was less variable. With lower N supply (N < 180 kg ha−1) under careful management, nitrogen use efficiency (NUE) for field‐grown tomato was ≈0.4 Mg fresh fruit (kg N)−1 and average crop N accumulation increased from 37 to 210 kg N ha−1 as N fertilization increased from 0 to 333 kg N ha−1. As a fraction of the fertilizer N applied N fertilizer recovery ranged from 0.36 to 0.74 and 0.61 to 0.96 for drip‐irrigated and subirrigated crops, respectively.
Greenhouse experiments were conducted to study the effect of plant growth promoting rhizobacteria (PGPR; Bacillus pumilus SE 34, Pseudomonas putida 89B61, BioYield, and Equity), acibenzolar-S-methyl (Actigard), and a soil amendment with S-H mixture (contains agricultural and industrial wastes such as bagasse, rice husk, oyster shell powder, urea, potassium nitrate, calcium super phosphate, and mineral ash) on bacterial wilt incidence caused by Ralstonia solanacearum (race 1, biovar 1) in susceptible tomato (Lycopersicon esculentum cv. Solar Set). In experiments with PGPR, Pseudomonas putida 89B61 significantly reduced bacterial wilt incidence when applied to the transplants at the time of seeding and 1 week prior to inoculation with Ralstonia solanacearum. BioYield, a formulated PGPR that contained two Bacillus strains, decreased disease significantly in three experiments. Equity, a formulation containing more than 40 different microbial strains, did not reduced wilt incidence compared with the untreated control. With inoculum at low pathogen densities of 1 × 105 and 1 × 106 CFU/ml, disease incidence of Actigard-treated plants was significantly less than with nontreated plants. This is the first report of Actigard-mediated reduction of bacterial wilt incidence in a susceptible tomato cultivar. When PGPR and Actigard applications were combined, Actigard plus P. putida 89B61 or BioYield reduced bacterial wilt incidence compared with the untreated control. Incorporation of S-H mixture into infested soil 2 weeks before transplanting reduced bacterial wilt incidence in one experiment. Combination of Actigard with the S-H mixture significantly reduced bacterial wilt incidence in tomato in two experiments.
Protection of crops from bacterial diseases presents a continuing challenge, mandating the development of novel agents and approaches. Photocatalysis is a process where chemically reactive oxygen species are catalytically generated by certain minerals in the presence of light. These reactive oxygen species have the capacity to destroy organic molecular structures critical to pathogen viability. In this study, the antibacterial potential of photocatalytic nanoscale titanium dioxide (TiO(2)), nanoscale TiO(2) doped (incorporation of other materials into the structure of TiO(2)) with silver (TiO(2)/Ag), and nanoscale TiO(2) doped with zinc (TiO(2)/Zn; AgriTitan) was evaluated against Xanthomonas perforans, the causal agent for bacterial spot disease of tomato. In vitro experiments on photocatalytic activity and dose dependency were conducted on glass cover slips coated with the nanoscale formulations by adding a known population of X. perforans strain Xp-F7 and illuminating the cover slips under a visible light source. TiO(2)/Ag and TiO(2)/Zn had high photocatalytic activity against X. perforans within 10 min of exposure to 3 × 10(4) lux. Greenhouse studies on naturally and artificially infected transplants treated with TiO(2)/Zn at ≈500 to 800 ppm significantly reduced bacterial spot severity compared with untreated and copper control. Protection was similar to the grower standard, copper + mancozeb. The use of TiO(2)/Zn at ≈500 to 800 ppm significantly reduced disease incidence in three of the four trials compared with untreated and copper control, and was comparable to or better than the grower standard. The treatments did not cause any adverse effects on tomato yield in any of the field trials.
Volatile plant essential oils thymol and palmarosa oil, used at a concentration of 0.7%, were evaluated under field conditions for control of bacterial wilt of tomato caused by Ralstonia solanacearum. The experimental fields were artificially infested with the bacterial pathogen. Two hours after infestation, the plant essential oils were applied, then the plots were sealed with plastic mulch for 3 or 6 days. Tomato seedlings were transplanted into the field 7 days later. In fall of 2002, 92.5% of tomato plants (cv. Equinox) wilted in the untreated control plots. Both thymol and palmarosa oil soil treatments reduced bacterial wilt incidence significantly. Thymol was more effective than palmarosa oil based on the final assessment, when 33.1 and 48.1% of the plants had wilted in plots treated with thymol and palmarosa oil, respectively. Soil treatment with either thymol or palmarosa oil produced significantly higher yield of tomato than the untreated control. In 2003, only thymol was evaluated. Thymol application significantly reduced bacterial wilt incidence on the susceptible cultivar Solar Set. Disease incidence in untreated plots reached 65.5%, while in thymol treated plots only 12% of plants wilted. Thymol treatment also increased yield of Solar Set significantly compared with the untreated control. This is the first report on the use of thymol for controlling a plant disease under field conditions, which indicated that this compound provided effective control of bacterial wilt on susceptible tomato cultivars when used as preplant treatment of soils. Because of its volatile property and broad-spectrum functions, thymol shows potential to be used as a soil biofumigant for the management of various plant pathogens.
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